Part:BBa_K4060100:Design
pBad ligated with RBS
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 125
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 65
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
In our Design 1, antitoxin MazE was chosen as the core effector component, with pBad promoter and RNA thermometer acting as sensor components, and the TetR gene and TetR repressible promoter working together as an inverting logic gate. If all goes as expected, MazE will express under 37°C if the medium is free of L-arabinose, but the expression level will drop significantly either with the presence of L-Arabinose or when the ambient temperature is low. To evaluate the behavior of our system, we created three constructs: Firstly, we put all parts together as our plan in Constriuct 1. In Construct 2 , we would like to visualize how MazE will express under different L-Arabinose concentration and ambient temperature for quick check and further assays, so we replaced MazE with RFP. By the same token, we replaced TetR with RFP in Construct 3 to evaluate the expression level of TetR under different concentrations of L-Arabinose in case we needed to troubleshoot the system.
Fig1. Design logic of our composite parts.
Source
The parts are from 2019 igem kits.